Secretomes of human pluripotent stem cell-derived smooth muscle cell progenitors upregulate extracellular matrix metabolism in the lower urinary tract and vagina

Guobing Zhuang,Qingchun Shao,Darlene Tran,Hongbo Wang,Bertha Chen,Yan Wen,Mason Briggs

doi:10.1186/s13287-021-02292-y

Abstract

BackgroundAdult mesenchymal stem cells (MSCs) have been studied extensively for regenerative medicine; however, they have limited proliferation in vitro, and the long culture time induces cell senescence. MSCs also contribute to tissue repair through their paracrine function. In this study, we sought to examine the paracrine effects of human smooth muscle cell progenitors (pSMC) on the urethra and adjacent vagina of stress urinary incontinence rodents. We use human pluripotent stem cell (PSC) lines to derive pSMCs to overcome the issue of decreased proliferation in tissue culture and to obtain a homogenous cell population.MethodThree human PSC lines were differentiated into pSMCs. The conditioned medium (CM) from pSMC culture, which contain pSMC secretomes, was harvested. To examine the effect of the CM on the extracellular matrix of the lower urinary tract, human bladder smooth muscle cells (bSMCs) and vaginal fibroblasts were treated with pSMC-CM in vitro. Stress urinary incontinence (SUI) was induced in rats by surgical injury of the urethra and adjacent vagina. SUI rats were treated with pSMC-CM and monitored for 5 weeks. Urethral pressure testing was performed prior to euthanasia, and tissues were harvested for PCR, Western blot, and histological staining. Kruskal-Wallis one-way ANOVA test and Student t test were used for statistical comparisons.ResultspSMC-CM upregulated MMP-2, TIMP-2, collagen, and elastin gene expression, and MMP-9 activity in the human bladder and vaginal cells consistent with elastin metabolism modulation. pSMC-CM treatment in the SUI rat improved urethral pressure (increase in leak point pressure compared to intact controls, p < 0.05) and increased collagen and elastin expression in the urethra and the adjacent vagina.ConclusionConditioned media from smooth muscle cell progenitors derived from human pluripotent stem cells improved urethral leak point pressure and collagen and elastin content in the SUI rat. These findings suggest a novel therapeutic potential for PSC-based treatments for SUI and pelvic floor disorders where tissues are affected by collagen, elastin, and smooth muscle loss.

Highlights

Adult mesenchymal stem cells (MSCs) have been studied extensively for regenerative medicine; they have limited proliferation in vitro, and the long culture time induces cell senescence
Results: Smooth muscle cell progenitors (pSMC)-conditioned medium (CM) upregulated Matrix metalloproteinase (MMP)-2, Tissue inhibitor of metalloproteinase (TIMP)-2, collagen, and elastin gene expression, and MMP-9 activity in the human bladder and vaginal cells consistent with elastin metabolism modulation. Smooth muscle cell progenitors conditioned media (pSMC-CM) treatment in the Stress urinary incontinence (SUI) rat improved urethral pressure and increased collagen and elastin expression in the urethra and the adjacent vagina
These findings suggest a novel therapeutic potential for pluripotent stem cell (PSC)-based treatments for SUI and pelvic floor disorders where tissues are affected by collagen, elastin, and smooth muscle loss

Summary

Introduction

Adult mesenchymal stem cells (MSCs) have been studied extensively for regenerative medicine; they have limited proliferation in vitro, and the long culture time induces cell senescence. We sought to examine the paracrine effects of human smooth muscle cell progenitors (pSMC) on the urethra and adjacent vagina of stress urinary incontinence rodents. We use human pluripotent stem cell (PSC) lines to derive pSMCs to overcome the issue of decreased proliferation in tissue culture and to obtain a homogenous cell population. Stress urinary incontinence (SUI) is one of the most prevalent pelvic floor disorders affecting many adults worldwide [1]. Current treatment options include pelvic floor physical therapy [7], bulking agent injections into the urethral tissue, or surgery with placement of synthetic mesh sling around the midurethra to provide support [8,9,10]. Surgical treatment is efficacious but the adverse events, including general risks of surgery, synthetic mesh complications, nerve injury, bladder perforation, postoperative voiding dysfunction [11, 12], and long-term recurrence rates, have led to a quest for the development of new approaches

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